Project Summary/Abstract: The goal of this project is to develop a new highly sensitive and cost effective Reverse Transcription Quantitative Bioluminescence Assay (RT-qBLA) for high-throughput detection and quantification of large (mRNA) and small (microRNA) RNAs in diverse biological samples. The RT-qBLA implements the same detection concept known from pyrosequencing, yet expanding pyrosequencing detection methodology for highly sensitive and accurate quantification of RNA molecules for gene expression analysis. The proposed assay can successfully challenge the real time quantitative RT-PCR technique (RT-qPCR) in many applications and can be superior to RT-qPCR particularly for the analysis of small RNA molecules. The RT-qBLA is requiring less expensive reagents and equipment and can be used in a number of commercial assays for applications in life sciences research, drug discovery, clinical diagnosis, environmental analysis, and biodefense. Project Narrative: The goal of this project is to develop a new high performance cost effective bioluminescence assay for gene expression analysis. The proposed assay provides technologically advanced and significantly less expensive alternative for real time polymerase chain reaction technology, which currently dominates in the market. The proposed bioluminescent assay is universal and can be used in various applications in life science research, drug discovery, clinical diagnosis and biodefense in more than 1,800 clinical laboratories and thousands of life science research laboratories in the U.S. [unreadable] [unreadable] [unreadable]